Dear all,
We are pleased to announce this year’s Proteored Multicentric Experiment, PME7, focused in Quantitative Proteomic Analysis. This year we count on the valuable collaboration of SIGMA-ALDRICH which as agreed to supply us samples of a “Quantitative Proteomics Sample Set” they have prepared and have been working on. Analysis of this sample set was presented at the last ASMS conference. The poster can be downloaded here. More details about the samples will be also provided through this web page: the protein identities, suggested chromatographic conditions, a table of suggested SRM transitions to be used in the analysis, etc. (see sample information section).
Samples and Objectives of PME7:
The sample set you will receive for analysis consists on 5 different samples A-E. Each one contains tryptic digests of 9 human proteins, spiked in different amounts to a yeast lysate digest. The amounts of these proteins are distributed in three different concentration “Tiers”:
Tier1: 3 protein digests in the range 25-2500 fmol / microgram of yeast lysate
Tier2: 3 protein digests in the range 2.5- 1250 fmol /microgram of yeast lysate
Tier3: 3 protein digests in the range 0.25-25 fmol / microgram of yeast lysate.
In addition, each sample contains identical amount (250 fmol) of a tenth protein digest, for normalization purposes.
Finally, two different labeled AQUA peptides per protein have been added to each sample in defined amounts.
You will be receive dried aliquots of samples A-E, containing each 15 microgram of yeast lysate digest plus the corresponding protein digests and AQUA peptides added, enough for several replicate analysis of each sample.
Altogether, this sample set provides a versatile system to evaluate quantitative analysis methods. Both relative quantitation (between the different samples) and absolute quantitation (referred to the labeled peptides present) can be assessed. The sample can be useful to evaluate analysis by different methods, such us iTRAQ, MSE, or MRM.
The samples will be sent as digests, ready for LC-MS analysis, so different tryptic digestion conditions are not going to be an issue. Replicate LC-MS analysis in multiple labs / using different approaches should allow interesting insights into many issues on quantitative proteomics: accuracy, sensitivity, across lab reproducibility, scope and limitations of each method, etc.
Calendar
The samples are ready to be distributed. We propose to send them during the last week of July.
To request the samples for the experiment, please send an e-mail to fcanals at vhio.net with the subject “PME7 sample request” and your mail address in the body of the message, as soon as possible.
The results of the analysis will be discussed during the next Proteored WG1-2 meeting, which will take place in La Cristalera, (Miraflores de la Sierra, Madrid) the 30th November - 1st December, 2011.
As in previous multicentric experiments, we will ask to submit your results for a centralized analysis, comparisons, etc. Instructions on results submission, templates to be used, MIAPE documents, etc will be detailed on the PME7 webpage. Deadline for results submission will be 15th November 2011.
Any comments, suggestions are welcome.
Best regards,
Francesc
On behalf of Proteored Working Group 1 and 2 |
